Plant Biology

1. Since plants interact with their environment through complex combinations of phytochemicals rather than metabolites in isolation, quantifying plant chemodiversity is of increasing interest. Although functionally relevant, structural disparity is mostly neglected in measures of chemodiversity because its integration relies on known compound identity, limiting broad application. 2. We established an approach deriving compound dissimilarity from UV-Vis spectra in HPLC-DAD datasets, evaluated how it relates to structure- or biosynthesis-based approaches, and examined whether it provides meaningful contributions to chemodiversity measures. For this, we applied it in an experiment full-factorially testing the effects of drought and herbivory on Solanum dulcamara leaf chemodiversity. 3. UV-Vis spectral dissimilarity aligned well with fMCS-based structural dissimilarity and reflected structural relationships within a set of standards. Incorporating it into chemodiversity analysis improved separation of the effects of drought and herbivory. Especially in the combination of both stresses, their distinct effects on different leaf metabolites were only fully reflected when accounting for compound disparity. 4. Hence, UV-Vis spectral dissimilarity captures chemically meaningful compound relatedness without requiring compound identity. Using it as a proxy for compound disparity adds a biologically relevant dimension to measures of chemodiversity with broader implications when assessing functional consequences of phytochemical diversity.

Fleshy fruits have evolved diverse traits to attract seed dispersers in response to frugivore behavior and sensory capacities. Fruit scent has been suggested to signal ripeness and nutritional quality, yet the volatile components involved and the information they convey remain poorly understood. It is unknown which information is encoded in fruit scent, whether plants actively synthesize these signals, and thus whether scent constitutes an evolved communication system shaping seed-dispersal interactions. Aliphatic esters, chemicals whose odor is often described as fruity, are abundant in some ripe fruits, especially those dispersed by animals which tend to rely on their sense of smell for fruit selection. Moreover, they have been argued to be associated with sugar content, potentially rendering them an honest signal and hence a hotspot of animal-plant chemical communication. We investigated whether aliphatic esters indicate fruit quality honestly and represent an adaptive trait shaped by disperser identity. Using 13 fig species (Ficus spp.; Moraceae) in Madagascar, we quantified seed dispersal by multiple animals using a quantitative ecological network. We then quantified chemical signals and nutritional rewards using thermal desorption gas chromatography-mass spectrometry (TD-GCMS) and high-performance liquid chromatography (HPLC), and used genome-guided transcriptome assembly to identify the candidate genes responsible for ester signaling. Our results show that (a) aliphatic esters occur more frequently in species dispersed primarily by olfactory-oriented mammals than in those dispersed by visually oriented birds; (b) ester abundance correlates positively with soluble sugar across species only in mammal-dispersed species, indicating an honest signal that is activated only when ecologically relevant; and (c) putative AAT gene revealing elevated expression associated with the increased abundance of aliphatic esters, sugars in single mammal-dispersed taxon. Together, these findings support the hypothesis that fruits have evolved to utilize the biochemical link between esters (signals) and sugars (rewards) to provide honest signals to seed dispersers.

Flower color has primarily been studied in the context of pollinator attraction, although effects on thermal energy balance are also important, especially in the context of global climate change. We used infrared imaging to compare petal temperatures of white versus pigmented cultivars of ten angiosperm taxa under controlled environmental conditions. Excised sets of flowers (n= 6 sets per species) exhibiting white, light, and/or dark anthocyanin (red to purple) coloration were mounted perpendicularly to the sun at mid-day, under clear sky, low wind (<1 m s-1) conditions. Sunlight was filtered through either UV-transparent or UV-opaque film, and petal temperatures were measured using an infrared camera after one minute equilibration. In all species, pigmented flowers were significantly warmer than lighter-colored conspecifics. Mean differences averaged +5.3{degrees}C for darker-colored versus white morphs, +2.9{degrees}C for lighter-colored versus white. Most warming was associated with visible wavelengths, but additional warming under UV-inclusion was also observed in some species. In situ observations of intact landscape plants under low-wind, high-light conditions corroborated experimental results, with differences exceeding 10{degrees}C observed in some taxa. Temperature differences >7{degrees}C were also recorded for purple versus white sections of the same flower in multicolored Viola and Petunia cultivars. Follow-up experiments using dark-pink and white varieties of Impatiens x hybrida corroborated well-known effects of sunlight intensity and wind speed on floral temperatures, helping to explain inconsistent reports in the literature. Our results clearly demonstrate that anthocyanin pigments can have significant and dramatic impacts on floral temperatures, which could be an important factor driving evolution of flower color. In the context of climate change, floral pigments could amplify the effects of rising global temperatures, negatively impacting plant reproduction and crop yields, especially on the warmer end of species ranges. Changes in flower color could also potentially induce shifts in pollinator communities, which could have community-scale effects.

Temperate forests increasingly face extreme air temperature, but plant physiological responses, particularly alterations in carbon allocation or protection via volatile organic compound (VOC) emissions, remain poorly understood. We pulse-labelled well-watered saplings of Fagus sylvatica and Pseudotsuga menziesii in a controlled heat stress experiment with 13CO2 to quantify heat-induced shifts in CO2, VOC and C pool exchange, specifically analyzing compound-specific {delta}13C of terpenoids, water-soluble organic matter (WSOM) and dark respiration. Under heat stress, up to 50% of fresh assimilates were directed to maintenance respiration and 1-2% to VOC emissions, while net assimilation and water use efficiency decreased by 50-75% in both species. Heat directly affected metabolic processes and reduced turnover rates of fresh assimilates in F. sylvatica, but accelerated them in P. menziesii. Strong 13C labelling of some compounds, particularly acyclic ones, suggested increased de novo synthesis of specific terpenoids for heat stress protection. By tracing the fate of recently assimilated 13CO2 we demonstrate that heat stress reduces net carbon uptake and water use efficiency, disrupts turnover of C pools and increases carbon loss via respiration and de novo synthesis of specific VOCs, potentially diminishing net carbon uptake of forests under future heat extremes.

Utricularia gibba L. is an aquatic carnivorous plant with a diverse set of capabilities. Reddening of traps frequently occurs in old in vitro cultures. While anthocyanins are often responsible for red coloration in plants, not every plant turns red. Stress factors like high light or excess sucrose have previously been shown to induce the formation of anthocyanins. Here, we hypothesized the red trap formation to be dependent on nutrient deprivation and tested nitrogen deprivation. The results suggest, that only in combination with light, nitrogen deficiency leads to the activation of the complete anthocyanin biosynthesis pathway and visible red coloration. However, in darkness, anthocyanin biosynthesis appears generally less active compared to light conditions and expression of most anthocyanin biosynthesis genes is not significantly upregulated under nitrogen deficiency.

Tomato production, yield and fruit quality face major challenges due to several factors, including the complex polygenic inheritance of agronomically relevant traits, biotic and abiotic stresses, and increasingly stringent regulations limiting the use of phytosanitary products. In this context, bioinoculants have emerged as a sustainable strategy capable of enhancing yield without compromising fruit quality, conferring protection against different stresses and exerting a minimal or no impact on environment and human health. In this study, we evaluated the effects and the underlying mechanisms by which Streptomyces sp. N2A, an actinobacteria isolated from soybean rhizosphere, promotes seed germination, vegetative growth and yield in tomato, without modifying fruit quality. The obtained results demonstrated that the bacterial treatment significantly improved seedlin[g]s emergence and growth and development in vegetative stage. At harvest, yield was also significantly enhanced, mainly driven by increased individual fruit weight, which was positively correlated with a thicker pericarp in fruits from N2A-treated plants. Transcriptional analysis during fruit development revealed a coordinated induction of auxin and cytokinin signaling pathways before and after anthesis, providing a hormonal framework that underlies the promotion of pericarp growth. This study provides evidence of the beneficial effect of inoculation with Streptomyces sp. N2A on tomato yield and constitutes the first report describing the modification of fruit morphology and expression of genes involved in phytohormonal modulation during early growth and development, induced by a plant growth-promoting Streptomyces.

Polyploidization can profoundly affect plant metabolite biosynthesis, yet its influence on chemodiversity remains poorly understood, despite the central role of chemodiversity in mediating plant interactions with the environment. The coexistence of facultative apomictic and sexual reproductive systems across ploidy levels in Hypericum provides an excellent model for investigating the evolution of chemodiversity following polyploidization. We analyzed ploidy levels and leaf metabolic fingerprints across selected populations of three Hypericum taxa, H. maculatum, H. perforatum subsp. perforatum and H. perforatum subsp. veronense. Polyploidization was common across all three taxa. Leaf metabolic fingerprints were more pronouncedly differentiated by the ploidy level of the mother plant (F0) than that of the offspring (F1). Although unique metabolic features emerged in plants of most ploidy levels, diploid plants exhibited fewer metabolic features than polyploid plants. Higher Shannon diversity, functional Hill diversity, and intensities of features belonging to specific chemical families were associated with higher F0 ploidy levels in H. perforatum subsp. perforatum, but not in H. maculatum and H. perforatum subsp. veronense. Our findings demonstrate that polyploidization can lead to rapid shifts in chemodiversity across generations in Hypericum. The fast divergence in chemodiversity associated with polyploidization in H. perforatum may contribute to its remarkable invasive potential.

European white oaks grow from the Mediterranean coast to Southern Scandinavia, a huge environmental gradient that has likely fostered environmental adaptation. In the face of climate change, leveraging adaptations through assisted gene flow could help improve drought tolerance and maintain forest health, but requires an understanding of the species-specific patterns of adaptation to be successful at the target location. In this study, three common gardens were established in Switzerland, Tuerkiye, and Austria for two European white oak species (Quercus robur, and Q. pubescens) using provenances from Central and Southern Europe. Almost 900 oak seedlings were measured at key water-use efficiency and life history traits for their first two year of life and genotyped with low coverage whole-genome sequencing. Trait heritability and environmental adaptation were then explored through pedigree-free animal models, while the genomic architecture of traits was mapped using a genome wide association study ("GWAS"). Across the species, the heritability of measured traits was moderate to high, but common garden had a strong impact, signalling an environmental effect on the phenotype. Adaptation to precipitation seasonality was detected in key productivity and growth traits for both species, but had a small effect on absolute trait values. The GWAS identified a striking 150 kbp association in the Cyclic Nucleotide-Gated Ion Channel gene family with leaf d13C values. This gene family is involved in stomata opening and likely impacts the intrinsic water use efficiency under stress. Together, the strong signals of phenotypic plasticity and rather weak signals of climatic adaptation in seedlings suggest that assisted gene flow in these two white oaks is relevant only for highly drought-sensitive populations, if conducted managers should focus on seeds sources with high precipitation seasonality and smaller leaf sizes.

White-barked birches extend to the northern limit of tree growth, and their bark is known to reduce solar damage during winter and early-spring by limiting solar heating and the incidence of harmful freeze-thaw events. The physical basis for this protection, however, has remained unclear. Here, we show that extracted betulin, the dominant triterpenoid responsible for the bark's whiteness, and Himalayan birch bark, both exhibit passive radiative cooling. Under low solar irradiance, bark and betulin-pellets reach temperatures below that of a shaded reference, and pellets cool more than bark. The cooling arises from high solar reflectance, which suppresses solar heating, and substantial mid-infrared emission, which drives radiative heat loss toward outer space. These findings help explain how bark-whiteness may contribute to protecting birch trees from solar-induced thermal stress.

Cannabis is well known for its pungent, skunk-like aroma. Recent chemical studies have identified prenylated and C6 volatile sulfur compounds as contributors to its skunky and citrus-like aromas, but the pathways that produce these compounds remain unknown. This gap limits efforts to explain variation in sulfur-aroma traits and to selectively enhance or reduce those traits. To address this gap, we used the known chemistry of sulfur-containing volatiles in Cannabis and characterized sulfur and volatile biosynthetic pathways in other plant species to select candidate enzyme groups. Because the GMO cultivar is anecdotally associated with a pronounced sulfurous aroma, reference protein sequences and profile hidden Markov models were used to search its version 1 (v1) primary high-confidence protein set of 55,790 sequences. These searches recovered 975 unique proteins. Sequence screening retained 941 candidates across 20 reporting categories; 939 contained all expected domains, while the two candidates assigned to the methionine gamma-lyase (MGL)-nearest category had no category-specific expected-domain rule. The largest reporting category comprised 359 proteins containing a cytochrome P450 domain, recovered through a search motivated by cytochrome P450 family 74 (CYP74) enzymes involved in oxylipin and plant volatile formation. Thirteen of these proteins were also recovered by at least one full-length CYP74 reference search. Other large reporting categories included 218 sugar-transferase, 83 glutathione-transferase, and 61 alcohol dehydrogenase candidates. Comparison with the Cannabis Expression Atlas linked 168 candidates to 128 annotated genes through 100%-identity amino-acid matches spanning at least 80% of each GMO v1 candidate protein. Twenty-nine genes were tissue-specific, including 13 root-specific and 6 trichome-specific genes. These results define candidates for biochemical testing and direct searches for additional enzymes acting upstream and downstream in Cannabis sulfur-volatile pathways.

Tropospheric ozone (O) is a major atmospheric pollutant that affects plant carbon metabolism, redox homeostasis, and secondary metabolism, including the biosynthesis and emission of biogenic volatile organic compounds (BVOCs). However, the contribution of BVOCs to O3 tolerance, particularly in tropical woody species, remains poorly understood. Here, we investigated whether acute O exposure (cumulative AOT40 of 3497.82 ppb h) induces alterations in photosynthetic performance, redox homeostasis, and BVOC partitioning in Eugenia uniflora. We evaluated gas exchange, photosynthetic pigments, ascorbate and glutathione pools, emitted BVOCs, modeled intercellular BVOC concentrations, and the relative carbon cost associated with BVOC emissions. O exposure significantly increased net CO2 assimilation without affecting stomatal conductance, transpiration, leaf water status, or chlorophyll concentrations, indicating maintenance of photosynthetic performance. Carotenoid concentrations and total glutathione decreased, whereas glutathione redox status was maintained. O induced marked compound-specific changes in BVOC composition and partitioning. Several monoterpenes appeared exclusively under O exposure, {gamma}-elemene emission increased significantly, and the relative distribution of individual BVOCs between the modeled intercellular and emitted pools was altered. These findings show that the response of E. uniflora to acute O exposure was characterized by interplay among carbon assimilation, glutathione redox regulation, and BVOC partitioning rather than by increased total volatile emission. Enhanced carbon assimilation occurred without additional carbon loss through BVOC release, while changes in the modeled intercellular pool indicate that part of the volatile response remained within the leaf. Our findings highlight BVOC partitioning as an important dimension of the plant response to oxidative stress and demonstrate that emission measurements alone may not fully capture the fate and potential physiological role of volatile carbon under O exposure. O_FIG O_LINKSMALLFIG WIDTH=178 HEIGHT=200 SRC="FIGDIR/small/743946v1_ufig1.gif" ALT="Figure 1"> View larger version (52K): org.highwire.dtl.DTLVardef@653af1org.highwire.dtl.DTLVardef@ca5forg.highwire.dtl.DTLVardef@1e641bforg.highwire.dtl.DTLVardef@1e68fae_HPS_FORMAT_FIGEXP M_FIG C_FIG BVOC Partitioning Contributes to Oxidative Stress Defence Under Acute O Exposure

{middle dot} Globally, forest ecosystems face widespread mortality events. However, the independent impacts of distinct stressors, such as heat stress vs edaphic drought, remain poorly understood and physiological early warning indicators for tree mortality are urgently required. {middle dot} We exposed well-watered saplings of Fagus sylvatica, Pseudotsuga menziesii and Picea abies to summer heat waves and subsequent natural winter-desiccation. Physiological parameters (e.g. gas exchange, water uptake velocity via 2H labelling, and volatile organic compound emissions) were monitored throughout the growing season and survival was assessed regularly until subsequent spring to capture immediate and delayed mortality as a consequence of legacy effects. {middle dot} Heat exposure without soil water deficit, followed by winter desiccation, triggered species-specific mortality rates (51.8% F. sylvatica, 48.2% P. abies, 16.9% P. menziesii), with P. abies exhibiting significantly faster mortality response than the other species. Reduced water uptake, lower stomatal conductance, impaired photosynthetic efficiency, and altered VOC emissions distinguished non-surviving from surviving saplings months before visible damage in all three species. {middle dot} Heat stress drives mortality independent of edaphic drought, with sub-lethal physiological indicators detectable up to 10 months before visual signs. These early warning indicators could enable damage detection before lethal thresholds are crossed, offering new strategies for mitigating climate change-driven forest decline.

Extrafloral nectaries (EFNs) are a widespread plant defense mutualism trait and are highly convergent, appearing in hundreds of plant lineages worldwide. Here we investigate a report of possible EFNs in Erythranthe angulosa, a recently-described California wildflower. We integrate field observations, insect bioassays, an induction experiment, and microscopy to test for signatures of EFN function, finding no evidence that the distinctive axillary swellings produced by E. angulosa function as EFNs. We also uncovered two distinct morphs at the type locality of E. angulosa that diverge in the number of axillary swellings produced, as well as other shoot architecture traits such as stem thickness, leaf size, and branch number. Although the axillary swellings appear to not function as EFNs, they remain a compelling morphological variant within the yellow monkeyflowers that may perform storage or another unknown function.

Photorespiration significantly influences terrestrial carbon fluxes, yet empirical measurements of its variability across tree species and temperature conditions remain limited, constraining predictions of vegetation and climate models. We quantified apparent photorespiratory CO2 loss (Lapp) and its temperature response for seven temperate broadleaf tree species in northern Europe, using in situ O2-shift measurements in Uppsala, Sweden during peak summer. Apparent loss was derived as the difference between net CO2 assimilation under ambient (Anet) and O2-free conditions at three leaf temperatures (25, 30, and 35 {degrees}C), spanning typical and heat-wave scenarios. Apparent photorespiratory CO2 loss showed pronounced interspecific variation and increased with temperature, while net photosynthesis remained relatively stable. The ratio of apparent loss to net photosynthesis ({phi} = Lapp/Anet) rose sharply with temperature, reaching species-mean values up to 0.94 at 35 {degrees}C, indicating that photorespiration can represent nearly the entirety of net carbon gain under heat stress even when leaves remain net CO2 sinks. Suppression of photorespiration under N2 and associated changes in leaf temperature systematically reallocated photosynthetic electron transport: the fraction of ambient electron transport rate (ETR) allocated to net CO2 assimilation declined with temperature, whereas the complementary fraction allocated to apparent photorespiratory loss and other O2-dependent sinks increased, with ETR-based apparent loss and its proportional expression rising steeply across the 25-35 {degrees}C range. Together, these in situ flux and partitioning measurements reveal high variability and strong temperature sensitivity in apparent photorespiration among temperate trees. Compared to crop-based parameterisations, the {phi} values we report for temperate trees are substantially higher and more temperature-dependent, providing species-specific constraints that can improve Farquhar-von Caemmerer-Berry-type vegetation model representations of photorespiration in forest ecosystems.

The increasingly severe droughts in the Amazon Basin make it urgent to understand the resilience of tropical tree species and their microbiota. Plant-associated fungi and bacteria (i.e. extended phenotype) modulate drought stress for their hosts, but their role in recovery dynamics remains poorly understood. To test the impact of different drought durations on the recovery of both physiology and microbiota of tropical trees, we followed the responses of nearly 1,000 seedlings belonging to seven tropical tree species of seasonally flooded (SF) forests in a greenhouse experiment. Seedlings were subjected to different droughts, reflecting a current, a projected and an extreme drought scenario of the French Guiana climate. Plant responses were monitored after the drought and after rewetting. Plant performance was estimated through leaf gas exchange, photochemical functioning, leaf water potentials and water-related traits as well as morphological traits. Bacterial and fungal leaf communities were characterized with respectively 16S and ITS2 markers using high-throughput sequencing. Increasing the duration of the drought reduced the ability of plants to recover physiological functions, with differences among species which were only partially predicted by their drought tolerance strategies. Bacterial diversity increased in most plant host species after mild drought but not under the most severe stress. Bacterial dispersion and turnover responses were strongly host species-specific, without a general directional pattern across species. Fungal communities showed greater compositional stability, but exhibited consistently higher turnover compared to bacterial communities during both drought and recovery, with no convergence toward control composition. Finally, none of the recovery networks mirrored the architecture of the control network, regardless of prior drought duration, demonstrating that the integrated extended phenotype does not recover even when individual traits show signs of recovery. Our results reveal that both physiological recovery and microbial community recovery are strongly shaped by the plant host species identity and drought duration This study widens knowledge of SF tropical forests, vulnerable habitats in the context of climate change, through the lens of the associated microbial communities and functional traits. Beyond the effects of an increasingly uncertain climate combined with a rise in the frequency of extreme events, our study places emphasis on including tree species extended phenotypes in considering their recovery dynamics.

Although the biosynthetic pathways of benzylisoquinoline alkaloids (BIAs) have been extensively investigated in several plant species, their transcriptional regulatory mechanisms remain only partially understood. Jasmonate (JA)-responsive group IX APETALA2/Ethylene Responsive Factor (AP2/ERF) transcription factors (TFs) are well-known regulators of specialized plant metabolism, including the biosynthesis of various alkaloids. However, their specific roles in BIA biosynthesis remain largely elusive. Here, we isolated five novel group IX AP2/ERF TFs, designated Benzylisoquinoline alkaloid Jasmonate-responsive AP2/ERF (BJE1-5), from Coptis japonica. Phylogenetic analysis revealed that Benzylisoquinoline alkaloid Jasmonate-responsive AP2/ERF (BJE) proteins belong to subclades distinct from group IXa, which contains well-known AP2/ERF TFs involved in alkaloid biosynthesis. Transient expression analyses in C. japonica protoplasts demonstrated that certain BJEs, particularly CjBJE3 and CjBJE5, positively regulated BIA biosynthetic genes through a mutual regulatory network among BJE members. Moreover, CjBJE3 expression was regulated by CjbHLH1, a unique-type basic helix-loop-helix (bHLH) TF specific to BIA-producing plants. Furthermore, heterologous expression of CjBJE3 and CjBJE5 in cultured Eschscholzia californica cells significantly enhanced the overall BIA production, particularly by increasing end-product benzophenanthridine BIAs, highlighting several uncharacterized biosynthetic genes clustered in the genome. Our findings suggest that BIA-producing species have developed a specific regulatory network comprised of CjbHLH1 and BJE TFs, providing valuable clues for identifying novel biosynthetic enzymes.

The ability of seeds to survive until dormancy recedes and the germination requirements are met is an adaptive strategy. Proteomics improves our understanding of the mechanisms that control the adaptation to environmental heterogeneity. In this study, we investigated two European beech populations from different habitats that differed in dormancy and germination traits. We found that the populations exhibited different germination strategies, which were reflected in coordinated but quantitatively different proteomic reprogramming. The Miekinia population exhibited stronger accumulation of proteins involved in nucleotide sugar biosynthesis, S-adenosylmethionine metabolism, and flavonoid biosynthesis. Enhanced nucleotide sugar biosynthesis indicates more intensive cell wall remodelling and carbohydrate metabolism, which support embryo growth and faster germination. Increased S-adenosylmethionine metabolism suggests the epigenetic and hormonal regulation of germination differences between populations. Higher flavonoid biosynthesis indicates an enhanced antioxidant capacity associated with environmental protection. In contrast, the Wisa population showed stronger accumulation of proteins involved in RNA processing, suggesting tighter post-transcriptional regulation and proteome reorganization during germination. Consistent with its deeper dormancy and later germination, the Wisa population appears to rely more on RNA-level regulation, whereas the Miekinia population prioritizes metabolic activation. These contrasting proteomic profiles likely reflect population-specific physiological strategies associated with dormancy depth and adaptation to different climatic conditions. HighlightProteomic reprogramming reveals population-specific germination strategies in European beech, linking dormancy depth with contrasting metabolic activation and RNA-level regulation during the transition from dormancy to germination.

Elevational gradients provide a framework for understanding how environmental filtering reorganises communities and interactions, but plant-pollinator interactions along temperate forest elevational gradients remain overlooked. We studied early-spring understorey communities at four forest sites spanning the foothills towards the timberline (450-1,000 m a.s.l.) in the Krkono[s]e Mountains, Czechia. Across six transects per elevation, we quantified flowering plant species richness, floral resources and traits, and video-recorded flowers, yielding 4,003 pollinator visits. We analysed elevational patterns in species richness, community composition, floral traits, and quantitative network characteristics. Visitation frequency and flowering plant and pollinator species richness peaked at intermediate elevations. The contribution of dipteran relative to hymenopteran pollinators increased towards higher elevations, principally because of non-hoverfly flies, whereas individual bee groups showed no uniform response. Floral resources and traits showed no uniform elevational responses, although total nectar sugar availability peaked at the highest site because of the dominant Vaccinium myrtillus. Most notably, both network-level specialisation and mean species-level specialisation were generally greater at the two higher elevations, whereas nestedness was lower and other network characteristics showed no consistent patterns. These findings suggest that shifts in pollinator composition and dominant floral resources potentially shaped interactions along the gradient. The increasing specialisation with elevation contrasts with the generalisation often expected under reduced partner availability and indicates that forest networks may follow elevational patterns not predicted from open habitats. Despite limited site-level replication, this study provides, to our knowledge, the first community-wide characterisation of plant-pollinator interactions along a temperate forest elevational gradient and identifies patterns requiring evaluation across replicated gradients.

Volatile organic compounds (VOCs) define the distinctive aroma of cannabis and critically influence consumer preference, cultivar authentication, and breeding programs. However, systematic characterization of VOC diversity across commercial drug-type cultivars remains limited. This study presents a comprehensive volatilomics-based phenotypic characterization of 165 commercial drug-type cannabis accessions using gas chromatography with flame ionization detection and mass spectrometry (GC-FID/MS). We identified 61 high-confidence VOCs assigned to three biosynthetic classes: terpenoids (n = 45), fatty acid-derived volatiles (n = 12) and amino acid-derived volatiles (n = 4), resolved into 12 subclasses. Analysis of variance revealed highly significant among-accession differences for all compounds (p < 0.001; 2 = 0.67-0.97), with repeatability estimates averaging 0.81 (range 0.50-0.95). Unsupervised clustering partitioned accessions into three distinct chemotypes (n = 90, 53, and 22), supported by principal component and t-SNE analyses. Machine learning-based feature selection identified a consensus panel of 12 discriminative compounds (camphene, -fenchene, sabinene, -terpinene, ({+/-})-limonene, -humulene, linalool, endo-fenchol, {Delta}3-carene, -thujene, {gamma}-terpinene and -phellandrene) that recovered the chemotype assignment of 32 of 33 held-out accessions. Estimated odor-activity screening ranked prenylthiol, -pinene, ({+/-})-limonene, linalool and myrcene highest among the odor-active compounds. All three chemotypes shared a prenylthiol-dominated core (67-77% of summed OAV) and were distinguished by the extent and nature of terpenoid modulation of that core: minimally modulated (Cluster ZERO), citrus-floral modulated (Cluster ONE) and pine-terpenic modulated (Cluster TWO). These findings indicate that volatile diversity in this panel can be summarized by three reproducible chemotypes, providing a quantitative basis for accession characterization and a foundation for future breeding and quality-assessment studies.

Sinia rhodoleuca, the sole species of the monotypic genus Sinia (Ochnaceae), was previously transferred to Sauvagesia based mainly on morphological similarities. However, its phylogenetic position has remained unresolved because molecular data for the species were unavailable. Here, we generated genomic data for Sinia rhodoleuca and reconstructed its phylogenetic position within Sauvagesieae. Our phylogenomic analyses consistently recovered Sinia rhodoleuca as sister to Indosinia, whereas the Neotropical Sauvagesia formed a distantly related lineage, rendering Sauvagesia broadly circumscribed non-monophyletic. Comparative morphological evidence further supports the close relationship between Sinia and Indosinia, particularly in their closely parallel secondary veins, lacerate stipules, and prominent petaloid staminodes, while differences in floral characters support their recognition as distinct genera. We therefore reinstate Sinia as a distinct genus and provide a revised taxonomic treatment of Sinia rhodoleuca. Our study demonstrates how phylogenomic evidence can resolve long-standing taxonomic uncertainties and reveal evolutionary relationships obscured by morphological similarity.